Traditional Chinese medicine composition for treating breast cancer complicated with anxiety disorder as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention discloses a traditional Chinese medicine composition for treating breast cancer complicated with anxiety disorder as well as a preparation method and application of the traditional Chinese medicine composition. The traditional Chinese medicine composition is mainly prepared from the following raw material medicines: astragalus membranaceus, herba epimedii, ligusticum wallichii, semen boitae, acanthopanax, rhizoma drynariae, selfheal, arisaema cum bile, turtle shell, nigrum fruit, salvia chinensis and artemisia apiacea. The invention further discloses a preparation method of the traditional Chinese medicine composition, a pharmaceutical preparation and application. Pharmacodynamic experiments prove that the traditional Chinese medicine composition can remarkably inhibit tumor growth of anxiety tumor-bearing mice and improve anxiety behavioral indexes, achieves dual treatment targets of tumor resistance and emotion regulation, and is particularly suitable for middle and advanced breast cancer combined anxiety patient groups needing long-term treatment.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and more specifically, to a traditional Chinese medicine composition for treating breast cancer combined with anxiety, and a preparation method and application thereof. Background Art

[0002] The latest data from the World Health Organization shows that in 2020, there were 2,261,419 new cases of breast cancer worldwide, making it the most common cancer in women. In addition to facing pressure from the disease itself, patients also have a higher incidence of psychological distress. It is reported that the incidence of anxiety is as high as 84%, which seriously affects the quality of life and prognosis of patients. Therefore, it is of great clinical value and social significance to treat anxiety while treating tumors. At present, Western medicine anti-anxiety treatment mainly includes two modes: drug therapy and psychotherapy. Western medicine anti-anxiety drug treatment (such as benzodiazepines The drug has adverse reactions such as respiratory inhibition and impact on daytime wakefulness quality, drug withdrawal rebound, addiction, etc., and there is a risk of metabolic conflict with tumor treatment drugs (such as competitive inhibition of CYP3A4 enzyme). Patients often have resistance and poor compliance. Western medicine psychological intervention treatment has a long cycle and high cost, and it is difficult to meet the needs of patients in the middle and late stages for rapid relief. Traditional Chinese medicine treatment has a long history and is more easily accepted by patients; and its multi-target and multi-level action characteristics have more advantages for breast cancer, a disease caused by multiple factors such as psychological, physiological and social factors, and it is worthy of widespread clinical application. Existing Chinese medicine compound prescriptions mostly focus on a single pathogenesis (such as soothing the liver and relieving depression or softening and dispersing nodules), and lack the formulation design of "cancer poison-qi stagnation-phlegm and blood stasis" synergistic intervention. Although the "Guidelines for the Diagnosis and Treatment of Breast Cancer with Integrated Chinese and Western Medicine" completed by the inventor in the early stage recommends that breast cancer with anxiety can be treated by soothing the liver and tonifying the kidney, there is no standardized Chinese medicine preparation on the market that is clearly targeted at breast cancer with anxiety. The clinic still relies on individualized decoctions, which have problems such as large fluctuations in efficacy and difficult quality control.

[0003] Therefore, for patients with breast cancer and anxiety disorder, it is necessary to develop a traditional Chinese medicine preparation that can achieve the dual therapeutic goals of anti-tumor and emotional regulation. Summary of the invention

[0004] In view of the above technical problems, one object of the present invention is to provide a new Chinese medicine composition, which can disperse stagnation and nourish the mind and effectively treat breast cancer complicated with anxiety.

[0005] Another object of the present invention is to provide a method for preparing the above-mentioned Chinese medicine composition.

[0006] Another object of the present invention is to provide a Chinese medicine preparation comprising the above-mentioned Chinese medicine composition and the use of the above-mentioned Chinese medicine composition and its pharmaceutical preparation in the preparation of a drug for breast cancer combined with anxiety.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating breast cancer complicated with anxiety disorder, wherein the traditional Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 15-30 parts of astragalus, 6-15 parts of epimedium, 6-15 parts of ligusticum chuanxiong, 9-15 parts of arborvitae, 12-18 parts of acanthopanax, 9-15 parts of drynaria, 9-15 parts of selfheal, 6-12 parts of cholerae, 9-24 parts of turtle shell, 12-18 parts of solanum nigrum fruit, 12-18 parts of lithospermum officinale, and 9-15 parts of artemisia annua.

[0009] The Chinese medicine formula of the present invention is based on the theory of "treating different diseases with the same medicine". It deeply analyzes the core compatibility rules of "Tiaoqi Yangshen Decoction" (regulating emotions) and "Li Chong Decoction" (eliminating symptoms and dispersing nodules) by the famous modern doctor Zhang Xichun. Through the strategies of "targeted enhancement" and "toxicity reduction and adaptation", a new compound prescription - Sanjie Yangshen Recipe is created, which preliminarily realizes the three-axis synergy of "strengthening the body - resolving phlegm and dispersing nodules - detoxifying and calming the mind", and can take into account tumors and anxiety at the same time. The whole prescription takes the pathological chain of "phlegm, blood stasis and toxicity - qi and blood malnutrition - disorder of spirit" as the hub, and constructs a linkage regulation network of viscera, qi and blood, and emotions. The specific synergistic mechanism is as follows:

[0010] (1) Strengthening the foundation to resolve the comorbidity of "cancer-anxiety"

[0011] Astragalus (the main drug) nourishes the spleen and lung qi, clears and removes toxins, enhances the body's immune surveillance function (such as promoting NK cell activity), and improves anxiety reactions under stress by regulating the hypothalamus-pituitary-adrenal axis. Secondly, astragalus is warm and ascending in nature. The liver belongs to wood and responds to the spring season. Its warmth and preference for regulation make it a good choice for nourishing the liver. Epimedium (the main drug) is bitter and slightly warm. It enters the foot Shaoyin kidney and foot Jueyin liver meridians. It nourishes tendons and strengthens bones, nourishes essence and blood, warms and nourishes the liver and kidneys, eliminates scrofula, cures paralysis, clears wind and improves eyesight, and benefits the mind and calms the spirit. It can intervene in the progression of hormone-sensitive breast cancer by regulating the balance of estrogen metabolism (inhibiting aromatase activity), and at the same time improve anxiety-related physical symptoms such as hot flashes and insomnia caused by hypothalamus-pituitary-gonadal axis dysfunction. Acanthopanax senticosus (minister drug) can replenish qi and strengthen the spleen. Its acanthopanax glycoside E can bidirectionally regulate central monoamine neurotransmitters (5-hydroxytryptamine and norepinephrine), relieving cancer-related fatigue while directly intervening in the neurobiological basis of anxiety. The three together replenish the "innate-acquired" foundation, repair the qi, blood, yin and yang that are damaged by tumors and emotions, and cut off the vicious cycle of "deficiency of vital energy → generation of phlegm and blood stasis → restlessness".

[0012] (2) Resolving phlegm, dispersing nodules and detoxifying, directly attacking the intertwined disease of "tangible evil and invisible stagnant fire"

[0013] Prunella vulgaris + Arisaema consanguineum (minister drug) work together to clear the liver and purge fire, resolve phlegm and dissipate nodules. Ursolic acid in Prunella vulgaris induces apoptosis of breast cancer cells, and Arisaema consanguineum dissipates phlegm and heat. The two not only reduce local tumors, but also relieve liver depression and fire-transforming anxiety by clearing the "phlegm and fire" evil. Solanum nigrum + Lithospermum officinale (minister drug) work together to detoxify and dissipate nodules. Solanine inhibits tumor angiogenesis, and Lithospermum officinale blocks the metastatic pathway. While inhibiting tumor proliferation, their bitter and cold nature can clear the "heat and toxicity disturbing the heart" syndrome caused by chemotherapy or cancer toxins. Vinegar turtle shell (adjuvant drug) nourishes yin and suppresses yang, softens and dissipates nodules. It reduces systemic inflammatory response by regulating the tumor microenvironment (such as inhibiting inflammatory factors such as IL-6), and indirectly improves the anxiety and depression induced by the disorder of the "inflammatory factor-brain-gut axis". The five together eliminate tumors and relieve heat and toxicity from the level of "phlegm, toxicity, and blood stasis", and simultaneously eliminate the pathological carriers of anxiety (phlegm and fire obscuring the orifices, toxicity and blood stasis disturbing the mind).

[0014] (3) Activating blood circulation, unblocking meridians and regulating the joints, connecting the interactive pathways of “local stasis-overall spiritual function”

[0015] Chuanxiong (adjuvant) promotes Qi and blood circulation. Its ligustrazine improves the hypoxic microenvironment of tumor tissue and inhibits VEGF expression. It also promotes the permeability of the blood-brain barrier and helps tranquilizing ingredients (such as cypress seeds) enter the brain to take effect. Drynaria fortunei (adjuvant) nourishes the kidney and promotes blood circulation. The flavonoids it contains promote bone matrix synthesis, relieve pain from bone metastasis, and block the negative feedback chain of "pain-anxiety-insomnia". Both of them are used for "unblocking", which not only breaks up local blood stasis and promotes the penetration of anti-tumor drugs, but also improves emotional symptoms through "smooth Qi and blood → tranquility of mind".

[0016] (4) Building a steady-state mechanism of “body and mind in harmony” by calming the mind and settling the spirit

[0017] Platycodon seed (the adjuvant drug) nourishes the heart and fluids, is rich in fat oil that moisturizes the intestines and promotes bowel movements, and harmonizes the dryness of the whole prescription. It improves anxiety-related intestinal flora disorders (such as constipation after chemotherapy that aggravates anxiety) by regulating "intestinal-brain interaction". Artemisia annua (the adjuvant drug) penetrates latent heat, clears internal fire, and guides the drug into the liver meridian. Artesunate not only inhibits the activity of tumor stem cells, but also downregulates excessive secretion of adrenocorticotropic hormone-releasing hormone in the hypothalamus, relieving stress-induced anxiety. The two work together, Platycodon seed calms the mind from the "heart", and Artemisia annua releases heat from the "liver". The two work together to regulate the pathogenesis of "excessive heart and liver fire", achieving a two-way synergy of anti-tumor and sedative hypnosis.

[0018] The whole formula forms a four-fold synergistic effect through the unique strategy of "tonifying deficiency to calm the mind and attacking toxins to calm the mind": (1) Immune-endocrine-neurotransmitter linkage: Astragalus regulates immunity, Epimedium balances hormones, and Acanthopanax stabilizes neurotransmitters, jointly preventing the interactive deterioration of "tumor progression-anxiety aggravation"; (2) Tumor reduction-mental stability mutual promotion: Prunella vulgaris, Arisaema consanguineum, Solanum nigrum fruit, and Lithospermum officinale directly inhibit tumors and eliminate the root cause of anxiety; Torreya grandis, as a tranquilizer, improves sleep and enhances tolerance to anti-cancer treatment; (3) Micro-anti-inflammatory-macro-regulating mind: Turtle shell inhibits inflammatory factors, and Artemisia annua regulates the hypothalamus-pituitary-adrenal axis, intervening in "inflammation-related anxiety" at the molecular level; (4) Local blood circulation-overall tranquilizing circulation: Chuanxiong and Drynaria fortunei improve tumor microcirculation and create conditions for the circulation of qi and blood for tranquilizing drugs. The whole formula embodies the TCM tumor treatment principle of "combining disease and syndrome, and treating body and mind together". While inhibiting the progression of breast cancer, it reshapes the patient's "spirit-form-qi" homeostasis through multi-system regulation, achieving the synergistic goal of "dispersing the nodules without forgetting to nourish the spirit, and calming the mind to help fight cancer".

[0019] Exemplarily, the Chinese medicine composition is mainly made of the following raw materials in parts by weight: 15 parts of Astragalus, 6 parts of Epimedium, 6 parts of Chuanxiong, 9 parts of Platycladus Semen, 12 parts of Acanthopanax, 9 parts of Drynaria, 9 parts of Prunella Vulgaris, 6 parts of Arisaema Cum Bile, 9 parts of Carapax Trichosanthis, 12 parts of Solanum nigrum fruit, 12 parts of Lithospermum officinale, and 9 parts of Artemisia annua.

[0020] Exemplarily, the Chinese medicine composition is mainly made of the following raw materials in parts by weight: 20 parts of Astragalus, 9 parts of Epimedium, 9 parts of Chuanxiong, 12 parts of Platycladus Semen, 15 parts of Acanthopanax, 12 parts of Drynaria, 12 parts of Prunella Vulgaris, 9 parts of Arisaema Cum Bile, 15 parts of Tricotide Carapace, 15 parts of Solanum nigrum fruit, 15 parts of Lithospermum officinale, and 12 parts of Artemisia annua.

[0021] Exemplarily, the Chinese medicine composition is mainly made of the following raw materials in parts by weight: 30 parts of Astragalus, 12 parts of Epimedium, 12 parts of Ligusticum chuanxiong, 15 parts of Platycladus orientalis, 18 parts of Acanthopanax senticosus, 15 parts of Drynariae Rhizoma, 15 parts of Prunella Vulgaris, 12 parts of Arisaema ciliata, 20 parts of Carapax Trichosanthis, 18 parts of Solanum nigrum fruit, 18 parts of Lithospermum officinale, and 15 parts of Artemisia annua.

[0022] The basic formula of each raw material medicine of the above-mentioned traditional Chinese medicine composition of the present invention can be further adjusted on the basis of the above-mentioned traditional Chinese medicine composition according to actual clinical symptoms to enhance the adaptability of the composition of the present invention and improve the therapeutic effect.

[0023] In a second aspect, the present invention provides a method for preparing the above-mentioned Chinese medicine composition for treating breast cancer combined with anxiety disorder, the preparation method comprising the following steps:

[0024] According to the formula, astragalus, epimedium, chuanxiong, cypress seed, acanthopanax, bone drynaria, selfheal, dandelion, turtle shell, solanum nigrum fruit, lycopodiella cava and artemisia annua are taken, and water is added for decoction. After decoction, the first filtrate is obtained by filtering, and the residue is retained;

[0025] Adding water to the medicinal residue and boiling it, filtering it after boiling to obtain a second filtrate;

[0026] Combine the two filtrates to obtain the product.

[0027] Furthermore, in the above preparation method, the amount of water added for each decoction is 10-15 times the total weight of the raw material medicine, and the amount of water added for the two decoctions can be the same or different; the time for each decoction is 30-90 minutes, and the time for each decoction can be the same or different.

[0028] Furthermore, in order to facilitate storage and preparation of various pharmaceutical dosage forms, the above preparation method may also include the step of concentrating the filtrate into a clear paste.

[0029] Furthermore, the clear paste has a relative density of 1.0-1.3 measured at 50°C-60°C.

[0030] In a third aspect, the present invention provides a traditional Chinese medicine preparation for treating breast cancer complicated with anxiety disorder, wherein the traditional Chinese medicine preparation uses the above-mentioned traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the above-mentioned preparation method as an active ingredient.

[0031] Furthermore, according to the actual dosage form requirements, the Chinese medicine preparation also includes pharmaceutically acceptable excipients, such as binders, disintegrants, lubricants, fillers, suspending agents, etc. According to conventional preparation processes, various preparations such as decoctions, granules, pills, capsules, tablets, powders, oral liquids, etc. are prepared.

[0032] The present invention also provides the use of the above-mentioned Chinese medicine composition or Chinese medicine preparation in preparing a medicine for treating breast cancer complicated with anxiety.

[0033] In addition, unless otherwise specified, the raw materials of the Chinese medicine composition of the present invention can be obtained commercially, and any range recorded in the present invention, including the end values ​​and any numerical value between the end values, and any sub-range of the Chinese medicine composition composed of the end values ​​or any numerical value between the end values, can achieve the purpose of treating breast cancer combined with anxiety.

[0034] The beneficial effects of the present invention are as follows:

[0035] The Chinese medicine composition of the present invention can effectively inhibit tumor growth in anxious tumor-bearing mice and improve behavioral indicators without adverse reactions, achieving the dual therapeutic goals of anti-tumor and emotional regulation, and filling the gap in therapeutic drugs for breast cancer combined with anxiety. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Shown are the changes in the body weight of mice during the anxiety-bearing tumor modeling period Note: Compared with the normal control group, *P<0.05, **P<0.005, ***P<0.001;

[0037] Figure 2 The results of elevated maze test after modeling of two groups of mice are shown; compared with the normal control group, *P<0.05, **P<0.005, ***P<0.001;

[0038] Figure 3 The elevated maze test trajectories of two groups of mice are shown; A represents normal control group mice, and B represents anxious tumor-bearing group mice;

[0039] Figure 4 The results of mouse tumor diameter measurement are shown; compared with the anxiety tumor model group, *P<0.05, **P<0.005, ***P<0.001;

[0040] Figure 5A , Figure 5B and Figure 5C Shows the results of in vivo imaging 2 weeks after inoculation of 4T1-LUC breast cancer cells;

[0041] Figure 6 Shown are the changes in body weight of mice in each group after tumor implantation intervention;

[0042] Figure 7 Shown are the elevated maze tests of mice in each group after intervention;

[0043] Figure 8 The tumor inhibition rate of mice after intervention is shown; IR (%) = (1-TWt / TWc) × 100%, where TWt is the tumor weight of the treatment group, and TWc is the tumor weight of the anxiety-bearing tumor model group. DETAILED DESCRIPTION

[0044] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0045] Example 1 A Chinese medicine composition for treating breast cancer combined with anxiety

[0046] Raw material formula: Astragalus 20g, Epimedium 9g, Chuanxiong 9g, Platycladus orientalis seed 12g, Acanthopanax 15g, Drynaria 12g, Prunella Vulgaris 12g, Arisaema cinerea 9g, Carapace of Trionycis 15g, Solanum nigrum fruit 15g, Lithospermum officinale 15g, Artemisia annua 12g.

[0047] Take each raw material drug and add water to decoct twice. For the first time, add water 10 times the total weight of the raw material drug, decoct for 40 minutes, filter, and obtain a filtrate. For the second time, take the drug residue and add water 10 times the total weight of the raw material drug, decoct for 30 minutes, filter, and combine the two filtrates to obtain.

[0048] Example 2 A Chinese medicine composition for treating breast cancer combined with anxiety

[0049] Raw material formula: Astragalus 15g, Epimedium 6g, Chuanxiong 6g, Platycladus orientalis seed 9g, Acanthopanax 12g, Drynaria 9g, Prunella Vulgaris 9g, Arisaema cinerea 6g, Carapace of Trionycis 9g, Solanum nigrum fruit 12g, Lithospermum officinale 12g, Artemisia annua 9g.

[0050] Take each raw material and boil it in water twice. For the first time, add water 12 times the total weight of the raw material, boil it for 90 minutes, filter and obtain a filtrate. For the second time, take the residue and add water 10 times the total weight of the raw material, boil it for 90 minutes, filter and combine the two filtrates, then concentrate it into a clear paste (the relative density measured at 50°C is 1.24-1.30), and dry it under reduced pressure to obtain the product.

[0051] Example 3 A Chinese medicine preparation (granules) for treating breast cancer combined with anxiety

[0052] Raw material formula: Astragalus 30g, Epimedium 12g, Chuanxiong 12g, Platycladus orientalis seed 15g, Acanthopanax 18g, Drynaria 15g, Prunella Vulgaris 15g, Arisaema cinerea 12g, Carapace of Trionycis 20g, Solanum nigrum fruit 18g, Lithospermum officinale 18g, Artemisia annua 15g.

[0053] Take each raw material and decoct it twice. For the first time, add water 12 times the total weight of the raw material, decoct for 60 minutes, filter and obtain a filtrate; for the second time, take the residue and add water 10 times the total weight of the raw material, decoct for 60 minutes, filter, combine the two filtrates, and then concentrate them into a clear paste (clear paste with a relative density of 1.24-1.30 measured at 50°C), dry under reduced pressure, and then make granules according to the granule process.

[0054] Example 4 Experimental study on the efficacy of the Chinese medicine composition of the present invention in treating breast cancer combined with anxiety

[0055] 1. Experimental Materials

[0056] Cells: 4T1-LUC (mouse breast cancer cell-LUC);

[0057] Animals: BALB / c, female, 4-6 weeks old, 132;

[0058] Drugs: Sanjie Yangshen Recipe (the Chinese medicine composition of Example 2 of the present invention), paclitaxel injection (30 mg: 5 ml, Sichuan Huiyu), escitalopram oxalate (10 mg×7, Sichuan Kelun), Pingxiao capsules (0.23 g×100, Xi'an Zhengda).

[0059] 2. Experimental Grouping

[0060] 1) Normal control group: 0.1 ml of normal saline was injected subcutaneously at the same site to simulate the breast pad grafting process;

[0061] 2) Anxiety tumor-bearing group: chronic restraint + injection of 5×10 5 Tumor cells.

[0062] 3. Animal Modeling

[0063] 3.1 Anxiety-induced tumor model

[0064] Adaptive feeding for 7 days. The anxiety model was prepared by chronic restraint stress method, with restraint for 4 hours per day (10:00-14:00) for 35 consecutive days. On the 28th day of chronic restraint stress modeling, 4T1-Luc cell line was inoculated into BALB / c mice, subcutaneously in the fourth pair of mammary pads on the left side of the abdominal wall, and chronic restraint stress was continued for 7 days after tumor implantation.

[0065] 3.2 Tumor inoculation

[0066] Take 4T1-LUC cells in the logarithmic growth phase, digest them with 0.25% trypsin and prepare a cell suspension of 5×106 / mL. Each mouse is inoculated with 0.1 ml of cell suspension and subcutaneously inoculated into the fourth pair of mammary pads on the left side of the abdominal wall. The mice are first anesthetized with ether and then injected with tumor cells. To ensure cell viability, the 4T1-LUC cell suspension is placed in a beaker containing ice cubes during the experiment, and the entire inoculation process is completed within 1 hour.

[0067] IV. Model Evaluation

[0068] Body weight, behavioral assessment, and tumor implantation success rate were used as evaluation indicators of the model.

[0069] 1) Weighing: The mice were weighed every 3 days during the baseline period (before modeling) and during the modeling period, and the body weight of the mice was recorded.

[0070] 2) Elevated maze test: 35 days after modeling, the mice were evaluated behaviorally using an elevated maze test to determine whether the anxiety modeling was successful.

[0071] 3) Tumor diameter measurement: After inoculation of 4T1-LUC breast cancer cells, the tumor growth at the inoculation site was observed daily, and the size of the tumor nodules was measured with a vernier caliper every 3 days. Tumor grafting success rate (%) = mice with tumor tissues grown / mice inoculated with tumor cells × 100%. The tumor grafting success rate was used to determine whether the tumor model was successful.

[0072] 4) In vivo imaging: When hard nodules were palpable in all tumor-bearing mice (one week after tumor inoculation), 5 mice were randomly selected from each group and the growth of tumor tissue was observed using small animal in vivo imaging equipment.

[0073] 5. Medication Treatment

[0074] After the model is successfully constructed, wait until the tumor volume of the tumor-bearing group grows to 75-100mm 3 At 1:00 pm, the drug administration started uniformly, and the administration volume was converted to 10 ml / kg. After the anxiety tumor-bearing group model was successfully established, it was randomly divided into two groups according to tumor diameter and body weight, n=12, and ear tags were given. For drug administration intervention, please refer to the experimental grouping.

[0075] 1) Normal control group: 0.1 ml of normal saline was injected subcutaneously at the same site to simulate the process of breast pad tumor grafting. After the second grouping, the animals were given drinking water by gavage, 0.2 ml / animal, once a day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / animal, twice a week, for 21 consecutive days.

[0076] 2) Anxiety tumor model group: chronic restraint + injection of 5×10 5 Tumor cells: After the second grouping, animals were given drinking water by gavage, 0.2 ml / animal, once a day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / animal, twice a week, for 21 consecutive days.

[0077] 3) Sanjie Yangshen prescription low-dose group: chronic restraint + injection 5×10 5 Tumor cells were grouped for the second time and given Sanjie Yangshen prescription 0.1729 g / mouse by oral gavage, 0.2 ml / mouse, once a day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / mouse, twice a week, for 21 consecutive days.

[0078] 4) Medium dose group of Sanjie Yangshen prescription: chronic restraint + injection of 5×10 5 Tumor cells were divided into groups for the second time. They were given Sanjie Yangshen prescription 0.3458 g / mouse by oral gavage, 0.2 ml / mouse, once a day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / mouse, twice a week, for 21 consecutive days.

[0079] 5) Sanjie Yangshen high-dose group: chronic restraint + injection 5×10 5Tumor cells were divided into groups for the second time. They were given Sanjie Yangshen prescription 0.6916 g / mouse by oral gavage, 0.2 ml / mouse, once a day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / mouse, twice a week, for 21 consecutive days.

[0080] 6) Chemotherapy group: chronic restraint + injection of 5×10 5 Tumor cells, paclitaxel 10 mg kg after the second grouping -1 , intraperitoneal injection, 2 times / week + oral gavage with drinking water, 0.2ml / mouse, once / day, for 21 consecutive days.

[0081] 7) Chemotherapy combined with citalopram group: chronic restraint + injection of 5×10 5 Tumor cells, after the second grouping, citalopram 10 mg kg -1 , gavage, once a day, for 21 consecutive days. Paclitaxel 10 mg kg was given once on Tuesday and Friday each week -1 , intraperitoneal injection, for 21 consecutive days.

[0082] 8) Sanjie Yangshen prescription low-dose combined chemotherapy group: chronic restraint + injection of 5×10 5 Tumor cells were divided into groups for the second time. After the second grouping, Sanjie Yangshen prescription was administered intragastrically at a rate of 0.1729 g / mouse. The dose was 0.2 ml / mouse. The dose was once a day for 21 consecutive days. Paclitaxel was administered at a rate of 10 mg / kg / mouse on Tuesdays and Fridays. -1 , intraperitoneal injection, for 21 consecutive days.

[0083] 9) The medium-dose Sanjie Yangshen prescription combined with chemotherapy group: chronic restraint + injection of 5×10 5 Tumor cells were divided into groups for the second time. After the second grouping, Sanjie Yangshen prescription was administered intragastrically at a rate of 0.3458 g / mouse. The dose was 0.2 ml / mouse. The dose was once a day for 21 consecutive days. Paclitaxel was also administered at a rate of 10 mg / kg / mouse on Tuesdays and Fridays. -1 , intraperitoneal injection, for 21 consecutive days.

[0084] 10) Sanjie Yangshen high-dose combined chemotherapy group: chronic restraint + injection of 5×10 5 Tumor cells were divided into groups for the second time. After the second grouping, Sanjie Yangshen prescription was administered intragastrically at a dose of 0.6916 g / mouse. The dose was 0.2 ml / mouse. The dose was once a day for 21 consecutive days. Paclitaxel was administered at a dose of 10 mg / kg / mouse on Tuesdays and Fridays. -1 , intraperitoneal injection, for 21 consecutive days.

[0085] 11) Pingxiao capsule group: chronic restraint + injection 5×10 5 Tumor cells, Pingxiao capsule 1000mg·kg after the second grouping -1 , oral gavage, 0.2 ml / mouse, once / day, for 21 consecutive days; combined with intraperitoneal injection of normal saline, 0.2 ml / mouse, twice / week, for 21 consecutive days.

[0086] 6. Efficacy Evaluation

[0087] After administration, the general condition of the mice was closely observed, including mental state, coat color, activity, diet, and urination and defecation. The injection site was observed every day for ulceration, redness, and swelling.

[0088] 1) Measurement of tumor diameter and volume: During the intervention period, mice were measured once every 3 days using a vernier caliper to record the tumor diameter and calculate the tumor volume. 3 )=ab 2 / 2 (a is the long diameter of the tumor, b is the short diameter of the tumor).

[0089] 2) Body weight monitoring: The changes in body weight of mice in each group were recorded during the baseline period (before intervention) and every 3 days during the intervention period, and the average body weight of mice in each group was calculated.

[0090] 3) Elevated maze test: After 21 days of intervention, an elevated maze test was used to conduct behavioral evaluation and assess the anxiety level of the animals.

[0091] 4) Tumor weight: After 21 days of intervention, all mice were killed, the tumor tissue under the mammary pad was peeled off, and the tumor inhibition rate was calculated after weighing using an electronic balance. Tumor inhibition rate = (1-TWt / TWc) × 100%, where TWt is the tumor weight of the treatment group, and TWc is the tumor weight of the anxiety tumor model group.

[0092] 8. Experimental Results

[0093] 1. Model evaluation

[0094] Construction of anxiety-bearing breast cancer mouse model: Chronic restraint stress modeling was established for 35 days, and tumor cells were injected on the 28th day of restraint.

[0095] (1) Weight changes: Before the anxiety-tumor model was established, there was no statistically significant difference in the baseline weight of the two groups of mice. During the chronic restraint stress modeling process, the weight of the mice in the normal group showed a gentle increase, while the weight of the mice in the anxiety-tumor model group fluctuated, first decreasing and then slowly increasing. Compared with the normal control group, their weight was generally lower. The data showed that from the 12th day of modeling, the weight of the mice in the anxiety-tumor model group was significantly lower than that of the mice in the normal control group, and the difference was statistically significant. Figure 1 .

[0096] (2) Behavioral assessment: Elevated maze experiment: Each mouse in the two groups was placed in the center of the maze with its head facing the open arm. The observer was at least 2 m away from the center of the maze, and the experiment was conducted in a separate experimental room to minimize human interference. The number of times the mouse entered the open arm and the closed arm during the experimental period (5 min) was recorded separately (the criteria for entering and exiting the arm were strictly defined, with all four limbs entering the arm or 80% of the animal's body entering the arm). Analysis indicators: The number of times the animal entered the open wall (open arm entry, OE) and the number of times the animal entered the closed wall (close arm entry, CE) were recorded, and the proportion of the number of times the animal entered the open wall was calculated (OE% = OE / (OE+CE) × 100%), which was used as an indicator for evaluating anxiety. 35 days after modeling, compared with the normal control group, the proportion of open arm times in the anxious tumor model group mice was significantly reduced, as shown in Figure 3. Figure 2 , Figure 3 The difference was statistically significant, P<0.05.

[0097] (3) On the 6th day after inoculation of 4T1-LUC cells, tumor nodules were palpable in all groups, and the tumor diameters were all greater than 2 mm × 2 mm. Figure 4 , the tumor implantation success rate was 100%. Two weeks after tumor implantation and before the second grouping, small animal live imaging detection was performed, and 5 mice were randomly selected from each group. Bioluminescence could be collected from each mouse, as shown in Figure 5. This proves that the anxious tumor-bearing breast cancer mouse model was successfully established.

[0098] 2. Efficacy evaluation

[0099] 2.1 Sanjie Yangshen prescription can improve the anxiety state of mice

[0100] (1) Weight changes

[0101] At the baseline stage, the body weights of mice in each modeling group were lower than those in the normal control group. During the intervention period, the body weights of mice in each modeling group fluctuated slightly, and the body weights of the chemotherapy combined with citrate group remained low. Compared with the normal control group, the differences between the groups were significant (P<0.05, see Figure 6 .

[0102] (2) Elevated maze test

[0103] After 21 days of intervention, the proportion of arm opening times in the medium-dose group of Sanjie Yangshen prescription was significantly increased compared with the anxious tumor model group (see Figure 7 ), P<0.05, which can improve anxiety. The proportion of arm opening times in the low-dose Sanjie Yangshen group was slightly increased, which can improve anxiety to a certain extent. Although the proportion of arm opening times in the low-dose Sanjie Yangshen group and the high-dose Sanjie Yangshen group was lower than that in the medium-dose group, it was better than the Pingxiao Capsule group. The low-dose Sanjie Yangshen group and the medium-dose Sanjie Yangshen group were significantly better than the same dose Sanjie Yangshen group combined with chemotherapy.

[0104] 2.2 Sanjie Yangshen Prescription can inhibit tumor growth

[0105] (1) According to tumor weight: After 21 days of intervention, the tumor tissue was removed and weighed. The tumor inhibition rate exceeded 40% in the medium-dose Sanjie Yangshen prescription group; the tumor inhibition rate exceeded 30% in the chemotherapy combined with citalopram group and the low-dose Sanjie Yangshen prescription combined with chemotherapy group. Sanjie Yangshen prescription can inhibit tumor growth and improve the efficacy of chemotherapy. Figure 8 .

[0106] (2) According to tumor volume, after 21 days of intervention, the tumor volumes of the chemotherapy combined with citalopram group, the Sanjie Yangshen low-dose combined with chemotherapy group, and the Sanjie Yangshen medium-dose combined with chemotherapy group were significantly lower than those of the model group, P < 0.05, and the tumor volume of the Sanjie Yangshen combined with chemotherapy group was slightly lower than that of the chemotherapy combined with citalopram group. This indicates that the Sanjie Yangshen formula can inhibit tumor growth and improve the efficacy of chemotherapy, and its effect is not inferior to that of chemotherapy drugs combined with anti-anxiety Western medicine. Figure 4 .

[0107] IX. Discussion

[0108] In summary, Sanjie Yangshen prescription, especially the medium-dose group, can improve patients' anxiety, inhibit tumor growth to a certain extent, improve the tumor inhibition effect of the chemotherapy group, and have a certain improvement on breast cancer complicated with anxiety disorders.

[0109] Example 5 Typical Cases

[0110] Zhu, female, 53 years old, was first diagnosed on March 5, 2023. She has been progression-free for more than 2 years and her anxiety has improved significantly.

[0111] More than 2 years after modified radical mastectomy for right breast cancer, after chemotherapy, luminal A type, T1N0M0

[0112] The patient underwent modified radical mastectomy for right breast cancer at Union Hospital on June 9, 2020. Postoperative pathology showed: (right breast) invasive ductal carcinoma, non-special type, vascular cancer thrombus (-), SLN (0 / 1), IHC: ER (+100%), PR (+15%), Her-2 (1+), ki-67 (10%). AC×4 was performed after surgery and letrozole is being treated. No genetic testing was performed.

[0113] At present: poor sleep (under anti-anxiety treatment), fatigue, sweating, dry throat, bitter taste in the mouth, good appetite, and normal bowel movements.

[0114] Previous experience: Underwent minimally invasive surgery for stage I rectal cancer in November 2018.

[0115] Auxiliary examination: February 6, 2023, biochemistry: TG: 2.52; February 28, 2023: right axillary lymph nodes enlarged, the largest 2.0×0.9cm. Hamilton anxiety score: 26 points, moderate anxiety.

[0116] Family history: maternal breast cancer.

[0117] Tongue and pulse: pale tongue with thin coating, stringy pulse.

[0118] prescription:

[0119] Astragalus 15g, Epimedium 6g, Chuanxiong 6g, Platycladus orientalis seed 9g

[0120] Acanthopanax senticosus 12g, Drynaria fortunei 9g, Prunella vulgaris 9g, Arisaema cinerea 6g

[0121] Turtle Shell 9g Solanum nigrum fruit 12g Lithospermum officinale 12g Artemisia annua 9g

[0122] White silkworm 15g light black bean 12g charred gardenia 9g turmeric 9g

[0123] ×14, one dose per day, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 5 days and rest 2 days.

[0124] Second visit: March 26, 2023

[0125] At present: cold feet, pale tongue with thin coating, stringy pulse. Hamilton Anxiety Score: 22 points, moderate anxiety.

[0126] Prescription: Prescription for the first diagnosis - light fermented black beans, + cinnamon 5g, Citrus aurantium 12g, × 14 packs, one dose per day, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 5 days and rest 2 days.

[0127] Third visit: May 17, 2023

[0128] At present: sleep is good, sweating is reduced, stomach bloating and acid reflux, lower limbs are cold, tongue is pale with thin coating, pulse is stringy. Hamilton Anxiety Score: 18 points, mild anxiety.

[0129] Prescription: Prescription for the second visit - Bombyx batryticatus, + Bletilla striata 15g, Salvia miltiorrhiza 15g, × 14 packs, one dose per day, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 6 days and rest 2 days.

[0130] Fourth diagnosis: July 2, 2023

[0131] At present: stomach bloating and acid reflux have improved, limbs are cold, tongue is pale with thin coating, pulse is stringy. Hamilton Anxiety Score: 15 points, mild anxiety.

[0132] Prescription: Prescription from the third visit - Bletilla striata, + light fermented black beans 12g, × 14 packs, one dose per day, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 6 days and rest 2 days.

[0133] Fifth visit: October 8, 2023

[0134] At present: limb coldness has improved, appetite, sleep and bowel movements are normal, tongue is pale with thin coating, pulse is stringy. Hamilton Anxiety Score: 12 points, possible anxiety.

[0135] Prescription: Four-diagnosis prescription - light fermented black beans, salvia miltiorrhiza, × 14 packs, one dose per 2 days, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 6 days and rest 2 days.

[0136] Sixth visit: February 25, 2024

[0137] At present: easy to wake up from sleep, normal bowel movements, vaginal bleeding once, pale tongue with thin coating, stringy pulse. Hamilton Anxiety Score: 9 points, possible anxiety.

[0138] Prescription: Five-diagnosis prescription + wolfberry 9g, × 7 packs, one dose per 2 days, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 6 days and rest 2 days.

[0139] Seventh visit: July 14, 2024

[0140] At present: sleep improved, no special discomfort, pale tongue coating, stringy pulse. Hamilton anxiety score: 6 points, no anxiety.

[0141] Prescription: Six-diagnosis prescription - Prunella vulgaris, + Prunella vulgaris 30g, Black Plum 12g, Cassia seed 30g, × 14 packs, one dose per 2 days, 2 times / day, 150ml / time, take half an hour after breakfast and dinner, take 6 and rest 2.

[0142] After that, the patient's anxiety symptoms improved significantly, and he continued to have regular outpatient follow-up visits, but there has been no progress so far.

[0143] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not limitations on the implementation methods of the present invention. For ordinary technicians in the relevant field, other different forms of changes or modifications can be made based on the above description. It is impossible to list all the implementation methods here. All obvious changes or modifications derived from the technical solution of the present invention are still within the protection scope of the present invention.

Claims

1. A Chinese medicine composition for treating breast cancer complicated with anxiety, characterized in that: The traditional Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 15-30 parts of astragalus, 6-15 parts of epimedium, 6-15 parts of ligusticum chuanxiong, 9-15 parts of cypress seeds, 12-18 parts of acanthopanax, 9-15 parts of drynaria, 9-15 parts of selfheal, 6-12 parts of cholerae, 9-24 parts of turtle shell, 12-18 parts of solanum nigrum fruit, 12-18 parts of lithospermum officinale, and 9-15 parts of artemisia annua.

2. The Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 15 parts of astragalus, 6 parts of epimedium, 6 parts of ligusticum chuanxiong, 9 parts of cypress seeds, 12 parts of acanthopanax senticosus, 9 parts of drynaria, 9 parts of selfheal, 6 parts of cholerae, 9 parts of turtle shell, 12 parts of solanum nigrum fruit, 12 parts of lithospermum officinale, and 9 parts of artemisia annua.

3. The Chinese medicine composition according to claim 1, characterized in that The Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 20 parts of astragalus, 9 parts of epimedium, 9 parts of ligusticum chuanxiong, 12 parts of cypress seeds, 15 parts of acanthopanax, 12 parts of drynaria, 12 parts of selfheal, 9 parts of cholerae, 15 parts of turtle shell, 15 parts of solanum nigrum fruit, 15 parts of lithospermum officinale, and 12 parts of artemisia annua.

4. The Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition is mainly prepared from the following raw materials in parts by weight: 30 parts of astragalus, 12 parts of epimedium, 12 parts of ligusticum chuanxiong, 15 parts of cypress seeds, 18 parts of acanthopanax, 15 parts of drynaria, 15 parts of selfheal, 12 parts of arisaema, 20 parts of turtle shell, 18 parts of solanum nigrum fruit, 18 parts of lithospermum officinale, and 15 parts of artemisia annua.

5. The method for preparing the Chinese medicine composition for treating breast cancer combined with anxiety according to any one of claims 1 to 4, characterized in that: The preparation method comprises the following steps: According to the formula, astragalus, epimedium, chuanxiong, cypress seed, acanthopanax, bone drynaria, selfheal, dandelion, turtle shell, solanum nigrum fruit, lycopodiella cava and artemisia annua are taken, and water is added for decoction. After decoction, the first filtrate is obtained by filtering, and the residue is retained; Adding water to the medicinal residue and boiling it, filtering it after boiling to obtain a second filtrate; Combine the two filtrates to obtain the product.

6. The preparation method according to claim 5, characterized in that: The amount of water added each time for decoction is 10-15 times the total weight of the raw material; The decoction time for each time is 30-90 minutes.

7. The preparation method according to claim 6, characterized in that: The preparation method further comprises the step of concentrating the filtrate into a clear paste; Preferably, the clear paste has a relative density of 1.0-1.3 measured at 50°C-60°C.

8. A Chinese medicine preparation for treating breast cancer complicated with anxiety, characterized in that: The Chinese medicine preparation uses the Chinese medicine composition according to any one of claims 1 to 4 or the Chinese medicine composition prepared by the preparation method according to any one of claims 5 to 7 as an active ingredient; Preferably, the traditional Chinese medicine preparation also includes pharmaceutically acceptable excipients.

9. The Chinese medicine preparation according to claim 8, characterized in that: The dosage form of the Chinese medicine preparation is decoction, granules, pills, capsules, tablets, powders or oral liquid.

10. Use of the Chinese medicine composition according to any one of claims 1 to 4 or the Chinese medicine preparation according to any one of claims 8 to 9 in the preparation of a medicament for treating breast cancer complicated with anxiety.

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